Regulation of the murine alpha A-crystallin promoter in transgenic mice.
To identify sequences necessary for lens-specific gene expression, lines of transgenic mice were generated which contain murine alpha A-crystallin promoter sequences [-111 to +46 (alpha 111), -88 to +46 (alpha 88), and -34 to +46 (alpha 34)] fused to the bacterial chloramphenicol acetyltransferase (CAT) gene and CAT expression was analyzed. Mice carrying the alpha 111-CAT or the alpha 88-CAT fusion transgene expressed CAT exclusively in lens, except for one line containing alpha 111-CAT, which expressed low levels of CAT in several nonlenticular tissues. Transcription from these promoters in lens initiated at the same site as the endogenous alpha A-crystallin promoter. In one line of mice alpha 88-CAT transgene became active in the lens during embryonic development at approximately the same time that the alpha A-crystallin gene normally begins to be expressed. In contrast, the alpha 34-CAT fusion transgene, containing the TATA box but no sequences further upstream, was inactive in transgenic mice. Our data suggest that 134 bp of sequence (-88 to +46) in the murine alpha A-crystallin gene is sufficient to provide lens specificity, although we cannot rule out the possibility that other sequences also contribute to promoter function.
Wawrousek
E F
EF
Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892.
Chepelinsky
A B
AB
McDermott
J B
JB
Piatigorsky
J
J
D000818:Animals; D001483:Base Sequence; D003459:Crystallins / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism*; D005314:Embryonic and Fetal Development; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation; D005796:Genes; D007908:Lens, Crystalline / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification
D003459:Crystallins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004247:DNA